Stabilized fibronectin domain compositions, methods and uses
First Claim
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1. A method of generating a protein scaffold binding to a specific target with a predefined binding affinity, comprising the steps of:
- a) contacting a library of a scaffold-based protein derived from a stability enhanced consensus sequence of a fibronectin type III domain incorporating randomized codons wherein a polynucleotide sequence encoding a polypeptide comprising a stability enhanced consensus sequence of a fibronectin type III domain of the amino acid sequence of SEQ ID NO;
150 is provided, randomizing codons are introduced into the polynucleotide sequence at selected positions and copies of the polynucleotide sequence are propagated to form a library of polynucleotides encoding variant scaffold proteins, with the specific target; and
b) isolating a protein scaffold binding to the specific target with the predefined affinity.
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Abstract
A protein scaffold based on a consensus sequence of fibronectin type III (FN3) proteins, such as the tenth FN3 repeat from human fibronectin (human Tenascin), including isolated nucleic acids that encode a protein scaffold, vectors, host cells, and methods of making and using thereof, exhibit enhanced thermal and chemical stability while presenting six modifiable loop domains which can be engineered to form a binding partner capable of binding to a target for applications in diagnostic and/or therapeutic compositions, methods and devices.
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Citations
4 Claims
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1. A method of generating a protein scaffold binding to a specific target with a predefined binding affinity, comprising the steps of:
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a) contacting a library of a scaffold-based protein derived from a stability enhanced consensus sequence of a fibronectin type III domain incorporating randomized codons wherein a polynucleotide sequence encoding a polypeptide comprising a stability enhanced consensus sequence of a fibronectin type III domain of the amino acid sequence of SEQ ID NO;
150 is provided, randomizing codons are introduced into the polynucleotide sequence at selected positions and copies of the polynucleotide sequence are propagated to form a library of polynucleotides encoding variant scaffold proteins, with the specific target; andb) isolating a protein scaffold binding to the specific target with the predefined affinity. - View Dependent Claims (2, 3, 4)
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Specification